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Pressure response of protein backbone structure. Pressure-induced amide 15N chemical shifts in BPTI

Published online by Cambridge University Press:  01 October 1999

KAZUYUKI AKASAKA
Affiliation:
Department of Molecular Science, The Graduate School of Science and Technology, Kobe University, 1-1 Rokkodai-cho, Nada-ku, Kobe 657-8501, Japan Department of Chemistry, Faculty of Science, Kobe University, 1-1 Rokkodai-cho, Nada-ku, Kobe 657-8501, Japan
HUA LI
Affiliation:
Department of Molecular Science, The Graduate School of Science and Technology, Kobe University, 1-1 Rokkodai-cho, Nada-ku, Kobe 657-8501, Japan
HIROAKI YAMADA
Affiliation:
Department of Chemistry, Faculty of Science, Kobe University, 1-1 Rokkodai-cho, Nada-ku, Kobe 657-8501, Japan
RENHAO LI
Affiliation:
Department of Biochemistry, University of Minnesota, 1479 Gortner Avenue, St. Paul, Minnesota 55108
TODD THORESEN
Affiliation:
Department of Biochemistry, University of Minnesota, 1479 Gortner Avenue, St. Paul, Minnesota 55108
CLARE K. WOODWARD
Affiliation:
Department of Biochemistry, University of Minnesota, 1479 Gortner Avenue, St. Paul, Minnesota 55108
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Abstract

The effect of pressure on amide 15N chemical shifts was studied in uniformly 15N-labeled basic pancreatic trypsin inhibitor (BPTI) in 90%1H2O/10%2H2O, pH 4.6, by 1H-15N heteronuclear correlation spectroscopy between 1 and 2,000 bar. Most 15N signals were low field shifted linearly and reversibly with pressure (0.468 ± 0.285 ppm/2 kbar), indicating that the entire polypeptide backbone structure is sensitive to pressure. A significant variation of shifts among different amide groups (0–1.5 ppm/2 kbar) indicates a heterogeneous response throughout within the three-dimensional structure of the protein. A tendency toward low field shifts is correlated with a decrease in hydrogen bond distance on the order of 0.03 Å/2 kbar for the bond between the amide nitrogen atom and the oxygen atom of either carbonyl or water. The variation of 15N shifts is considered to reflect site-specific changes in φ, ψ angles. For β-sheet residues, a decrease in ψ angles by 1–2°/2 kbar is estimated. On average, shifts are larger for helical and loop regions (0.553 ± 0.343 and 0.519 ± 0.261 ppm/2 kbar, respectively) than for β-sheet (0.295 ± 0.195 ppm/2 kbar), suggesting that the pressure-induced structural changes (local compressibilities) are larger in helical and loop regions than in β-sheet. Because compressibility is correlated with volume fluctuation, the result is taken to indicate that the volume fluctuation is larger in helical and loop regions than in β-sheet. An important aspect of the volume fluctuation inferred from pressure shifts is that they include motions in slower time ranges (less than milliseconds) in which many biological processes may take place.

Type
Research Article
Copyright
© 1999 The Protein Society

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